Objective To investigate the chemical compositions of Maxing Shigan Decoction(麻杏石甘汤,MXSGD)and elucidate its anti-influenza A virus(IAV)mechanism from prediction to validation.Methods Ultra high-performance liquid...Objective To investigate the chemical compositions of Maxing Shigan Decoction(麻杏石甘汤,MXSGD)and elucidate its anti-influenza A virus(IAV)mechanism from prediction to validation.Methods Ultra high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was employed to analyze the chemical compositions of MXSGD.Network pharmacology theories were used to screen and identify shared targets of both the potential targets of active ingredients of MXSGD and IAV.A protein-protein interaction(PPI)network was then constructed,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The binding stability between core bioactive compounds and key targets was validated by molecular docking and dynamic simulations.A total of 24 BALB/c mice were infected with IAV to build IAV mouse models.After successful modelling,the mouse models were randomly divided into model,MXSGD high-dose(2.8 g/kg),MXSGD low-dose(1.4 g/kg),and oseltamivir(20.14 mg/kg)groups,with an additional normal mice as control group(n=6 per group).The treatments were administered by gavage daily between 8:00 a.m.and 10:00 a.m.for five consecutive days.Upon completion of the administration,the body weight ratio,lung index,protein content in the bronchoalveolar lavage fluid(BALF),and the levels of inflammatory factors including interleukin(IL)-6 and tumor necrosis factor(TNF)-αin mice were measured to preliminarily analyze the therapeutic efficacy of MXSGD against IAV infection.Furthermore,the expression levels of mechanistic target of rapamycin(mTOR),hypoxia inducible factor(HIF)-1α,and vascular endothelial growth factor(VEGF)proteins in the HIF-1 signaling pathway,which was enriched by network pharmacology,were detected by Western blot.Results A total of 212 chemical components in MXSGD were identified by the UPLC-MS/MS method.These chemical components can be classified into 9 primary categories and 31 secondary categories.After intersecting the chemical component targets with IAV-related targets,a total of 567 potential MXSGD components targeting IAV were identified.The construction of PPI network and the results of both GO and KEGG enrichment analyses revealed that the anti-IAV effects of MXSGD were associated with multiple pathways,including apoptosis,TNF,HIF-1,and IL-17 signaling pathways.The results of molecular docking demonstrated that the binding energies between the core compound 1-methoxyphaseollin and key targets including HIF-1α,mTOR,and VEGF were all lower than–5.0 kcal/mol.Furthermore,molecular dynamics simulations confirmed the structural stability of the resulting complexes.Animal experiments showed that compared with the normal controls,IAV-infected mice showed significantly reduced body weight ratio,markedly increased lung index,protein content in BALF,and the levels of inflammatory factors such as IL-6 and TNF-α(P<0.01),thereby causing damage to the lung tissue;consequently,the expression levels of mTOR,HIF-1α,and VEGF proteins in the lung tissues of these mice were significantly elevated(P<0.01).However,after MXSGD treatment,the mouse models presented a significant increase in body weight ratio,as well as marked decreases in lung index,protein content in BALF,and the levels of inflammatory factors including IL-6 and TNF-α(P<0.01).Furthermore,the therapy alleviated IAV-induced injuries and significantly downregulated the expression levels of mTOR,HIF-1α,and VEGF proteins in lung tissues(P<0.01 or P<0.05).Conclusion MXSGD exerts anti-IAV effects through multi-component,multi-target,and multi-pathway synergism.Among them,1-methoxyphaseollin is identified as a potential key component,which alleviates virus-induced lung injury and inflammatory response via the regulation of HIF-1 signaling pathway,providing experimental evidence for the clinical application of MXSGD.展开更多
Influenza is an acute viral respiratory infection that has caused high morbidity and mortality worldwide.Influenza A virus(IAV)has been found to activate multiple programmed cell death pathways,including ferroptosis.F...Influenza is an acute viral respiratory infection that has caused high morbidity and mortality worldwide.Influenza A virus(IAV)has been found to activate multiple programmed cell death pathways,including ferroptosis.Ferroptosis is a novel form of programmed cell death in which the accumulation of intracellular iron promotes lipid peroxidation,leading to cell death.However,little is known about how influenza viruses induce ferroptosis in the host cells.In this study,based on network pharmacology,we predicted the mechanism of action of Maxing Shigan decoction(MXSGD)in IAV-induced ferroptosis,and found that this process was related to biological processes,cellular components,molecular function and multiple signaling pathways,where the hypoxia inducible factor-1(HIF-1)signaling pathway plays a significant role.Subsequently,we constructed the mouse lung epithelial(MLE-12)cell model by IAV-infected in vitro cell experiments,and revealed that IAV infection induced cellular ferroptosis that was characterized by mitochondrial damage,increased reactive oxygen species(ROS)release,increased total iron and iron ion contents,decreased expression of ferroptosis marker gene recombinant glutathione peroxidase 4(GPX4),increased expression of acyl-CoA synthetase long chain family member 4(ACSL4),and enhanced activation of hypoxia inducible factor-1α(HIF-1α),induced nitric oxide synthase(iNOS)and vascular endothelial growth factor(VEGF)in the HIF-1 signaling pathway.Treatment with MXSGD effectively reduced intracellular viral load,while reducing ROS,total iron and ferrous ion contents,repairing mitochondrial results and inhibiting the expression of cellular ferroptosis and the HIF-1 signaling pathway.Finally,based on animal experiments,it was found that MXSGD effectively alleviated pulmonary congestion,edema and inflammation in IAV-infected mice,and inhibited the expression of ferroptosis-related protein and the HIF-1 signaling pathway in lung tissues.展开更多
Objective To establish an objective method for evaluating the intrinsic characteristics between cold and hot nature of Chinese materia medica (CMM) through the different effects of Mahuang decoction (MHD) and Maxing S...Objective To establish an objective method for evaluating the intrinsic characteristics between cold and hot nature of Chinese materia medica (CMM) through the different effects of Mahuang decoction (MHD) and Maxing Shigan decoction (MSD) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of the temperature tropism between two groups of mice treated by MHD and MSD, respectively. Meanwhile, the activities of adenosine triphosphatase (ATPase), superoxide dismutase, succinate dehydrogenase, and malondialdehyde were measured. Results After treated by MHD, the macroscopic behavioral index of remaining rate on warm pad (40 ℃ ) of mice decreased significantly (P < 0.05), suggesting the enhancement of cold tropism, meanwhile, the internal indices of ATPase activity and oxygen consuming volume increased significantly (P < 0.05), suggesting the enhancement of energy metabolism. On the other hand, the above-mentioned indices in MSD group changed on the inverse way. Conclusion The relative drug nature of MHD and MSD revealed in this study is consistent with the theoretical prognostication or definition. It indicates that the internal cold and hot nature of CMM could be reflected in ethological way on the changes of animal temperature tropism which might be internally regulated by body energy metabolism.展开更多
Background:The COVID-19 has a huge negative impact on people’s health.Traditional Chinese Medicine(TCM)has a good effect on viral pneumonia.It is of great practical significance to study its pharmacology.Methods:The ...Background:The COVID-19 has a huge negative impact on people’s health.Traditional Chinese Medicine(TCM)has a good effect on viral pneumonia.It is of great practical significance to study its pharmacology.Methods:The ingredients and targets of each herb in Maxing Shigan Decoction which obtained from Traditional Chinese Medicine Systems Pharmacology(TCMSP)database,and the related targets of COVID-19 were screened by GeneCards database based on the network pharmacology.Venn was used to analyze the intersection target between active ingredients and diseases.Cytoscape software was used to construct an active ingredient-disease target network.The protein-protein interaction network was constructed by STRING database and Cytohubba was used to screen out the key targets.Gene Ontology(GO)functional enrichment analysis and KEGG pathway analysis were performed by DAVID database.Results:In this study,a total of 134 active ingredients and 229 related targets,198 targets of COVID-19 and 48 common targets of drug-disease were chosen.Enrichment items and pathways were obtained through GO and KEGG pathway analysis.The predicted active ingredients were quercetin,kaempferol,luteolin,naringenin,glycyrol,and the key targets involved IL6,MAPK3,MAPK8,CASP3,IL10,etc.The results showed that the active ingredients of Maxing Shigan Decoction acted on multiple targets which played roles in the treatment of COVID-19 by regulating inflammation,immune system and other pathways.Conclusions:The main contribution of this paper is to use data to mine the principles of the treatment of COVID-19 from the pharmacology of these prescriptions,and the results can be provided theoretical reference for medical workers.展开更多
基金Natural Science Foundation of Hunan Province(2025JJ80078)Open Fund of Hunan University of Chinese Medicine(21PTKF1005)。
文摘Objective To investigate the chemical compositions of Maxing Shigan Decoction(麻杏石甘汤,MXSGD)and elucidate its anti-influenza A virus(IAV)mechanism from prediction to validation.Methods Ultra high-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was employed to analyze the chemical compositions of MXSGD.Network pharmacology theories were used to screen and identify shared targets of both the potential targets of active ingredients of MXSGD and IAV.A protein-protein interaction(PPI)network was then constructed,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The binding stability between core bioactive compounds and key targets was validated by molecular docking and dynamic simulations.A total of 24 BALB/c mice were infected with IAV to build IAV mouse models.After successful modelling,the mouse models were randomly divided into model,MXSGD high-dose(2.8 g/kg),MXSGD low-dose(1.4 g/kg),and oseltamivir(20.14 mg/kg)groups,with an additional normal mice as control group(n=6 per group).The treatments were administered by gavage daily between 8:00 a.m.and 10:00 a.m.for five consecutive days.Upon completion of the administration,the body weight ratio,lung index,protein content in the bronchoalveolar lavage fluid(BALF),and the levels of inflammatory factors including interleukin(IL)-6 and tumor necrosis factor(TNF)-αin mice were measured to preliminarily analyze the therapeutic efficacy of MXSGD against IAV infection.Furthermore,the expression levels of mechanistic target of rapamycin(mTOR),hypoxia inducible factor(HIF)-1α,and vascular endothelial growth factor(VEGF)proteins in the HIF-1 signaling pathway,which was enriched by network pharmacology,were detected by Western blot.Results A total of 212 chemical components in MXSGD were identified by the UPLC-MS/MS method.These chemical components can be classified into 9 primary categories and 31 secondary categories.After intersecting the chemical component targets with IAV-related targets,a total of 567 potential MXSGD components targeting IAV were identified.The construction of PPI network and the results of both GO and KEGG enrichment analyses revealed that the anti-IAV effects of MXSGD were associated with multiple pathways,including apoptosis,TNF,HIF-1,and IL-17 signaling pathways.The results of molecular docking demonstrated that the binding energies between the core compound 1-methoxyphaseollin and key targets including HIF-1α,mTOR,and VEGF were all lower than–5.0 kcal/mol.Furthermore,molecular dynamics simulations confirmed the structural stability of the resulting complexes.Animal experiments showed that compared with the normal controls,IAV-infected mice showed significantly reduced body weight ratio,markedly increased lung index,protein content in BALF,and the levels of inflammatory factors such as IL-6 and TNF-α(P<0.01),thereby causing damage to the lung tissue;consequently,the expression levels of mTOR,HIF-1α,and VEGF proteins in the lung tissues of these mice were significantly elevated(P<0.01).However,after MXSGD treatment,the mouse models presented a significant increase in body weight ratio,as well as marked decreases in lung index,protein content in BALF,and the levels of inflammatory factors including IL-6 and TNF-α(P<0.01).Furthermore,the therapy alleviated IAV-induced injuries and significantly downregulated the expression levels of mTOR,HIF-1α,and VEGF proteins in lung tissues(P<0.01 or P<0.05).Conclusion MXSGD exerts anti-IAV effects through multi-component,multi-target,and multi-pathway synergism.Among them,1-methoxyphaseollin is identified as a potential key component,which alleviates virus-induced lung injury and inflammatory response via the regulation of HIF-1 signaling pathway,providing experimental evidence for the clinical application of MXSGD.
基金supported by the National Natural Science Foundation of China(No.81973670)the Natural Science Foundation of Hunan Province(No.2020J5418)Hunan Provincial Open Fund of the Key Laboratory of the Pathogen Biology of Integrated Traditional Chinese and Western Medicine(No.2022-KFJJ02).
文摘Influenza is an acute viral respiratory infection that has caused high morbidity and mortality worldwide.Influenza A virus(IAV)has been found to activate multiple programmed cell death pathways,including ferroptosis.Ferroptosis is a novel form of programmed cell death in which the accumulation of intracellular iron promotes lipid peroxidation,leading to cell death.However,little is known about how influenza viruses induce ferroptosis in the host cells.In this study,based on network pharmacology,we predicted the mechanism of action of Maxing Shigan decoction(MXSGD)in IAV-induced ferroptosis,and found that this process was related to biological processes,cellular components,molecular function and multiple signaling pathways,where the hypoxia inducible factor-1(HIF-1)signaling pathway plays a significant role.Subsequently,we constructed the mouse lung epithelial(MLE-12)cell model by IAV-infected in vitro cell experiments,and revealed that IAV infection induced cellular ferroptosis that was characterized by mitochondrial damage,increased reactive oxygen species(ROS)release,increased total iron and iron ion contents,decreased expression of ferroptosis marker gene recombinant glutathione peroxidase 4(GPX4),increased expression of acyl-CoA synthetase long chain family member 4(ACSL4),and enhanced activation of hypoxia inducible factor-1α(HIF-1α),induced nitric oxide synthase(iNOS)and vascular endothelial growth factor(VEGF)in the HIF-1 signaling pathway.Treatment with MXSGD effectively reduced intracellular viral load,while reducing ROS,total iron and ferrous ion contents,repairing mitochondrial results and inhibiting the expression of cellular ferroptosis and the HIF-1 signaling pathway.Finally,based on animal experiments,it was found that MXSGD effectively alleviated pulmonary congestion,edema and inflammation in IAV-infected mice,and inhibited the expression of ferroptosis-related protein and the HIF-1 signaling pathway in lung tissues.
基金National Basic Research Program of China ("973" Program) (2007CB5126072006CB504703)National Science Fund for Distinguished Young Scholars (30625042)
文摘Objective To establish an objective method for evaluating the intrinsic characteristics between cold and hot nature of Chinese materia medica (CMM) through the different effects of Mahuang decoction (MHD) and Maxing Shigan decoction (MSD) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of the temperature tropism between two groups of mice treated by MHD and MSD, respectively. Meanwhile, the activities of adenosine triphosphatase (ATPase), superoxide dismutase, succinate dehydrogenase, and malondialdehyde were measured. Results After treated by MHD, the macroscopic behavioral index of remaining rate on warm pad (40 ℃ ) of mice decreased significantly (P < 0.05), suggesting the enhancement of cold tropism, meanwhile, the internal indices of ATPase activity and oxygen consuming volume increased significantly (P < 0.05), suggesting the enhancement of energy metabolism. On the other hand, the above-mentioned indices in MSD group changed on the inverse way. Conclusion The relative drug nature of MHD and MSD revealed in this study is consistent with the theoretical prognostication or definition. It indicates that the internal cold and hot nature of CMM could be reflected in ethological way on the changes of animal temperature tropism which might be internally regulated by body energy metabolism.
基金the National Natural Science Foundation of China(Nos.62072157 and 61802116)the Natural Science Foundation of Henan province(202300410102)+1 种基金the Doctoral program of Henan Institute of Technology(KQ2002)the Science and Technology Research Key Project of Henan Province(No.192102210113).
文摘Background:The COVID-19 has a huge negative impact on people’s health.Traditional Chinese Medicine(TCM)has a good effect on viral pneumonia.It is of great practical significance to study its pharmacology.Methods:The ingredients and targets of each herb in Maxing Shigan Decoction which obtained from Traditional Chinese Medicine Systems Pharmacology(TCMSP)database,and the related targets of COVID-19 were screened by GeneCards database based on the network pharmacology.Venn was used to analyze the intersection target between active ingredients and diseases.Cytoscape software was used to construct an active ingredient-disease target network.The protein-protein interaction network was constructed by STRING database and Cytohubba was used to screen out the key targets.Gene Ontology(GO)functional enrichment analysis and KEGG pathway analysis were performed by DAVID database.Results:In this study,a total of 134 active ingredients and 229 related targets,198 targets of COVID-19 and 48 common targets of drug-disease were chosen.Enrichment items and pathways were obtained through GO and KEGG pathway analysis.The predicted active ingredients were quercetin,kaempferol,luteolin,naringenin,glycyrol,and the key targets involved IL6,MAPK3,MAPK8,CASP3,IL10,etc.The results showed that the active ingredients of Maxing Shigan Decoction acted on multiple targets which played roles in the treatment of COVID-19 by regulating inflammation,immune system and other pathways.Conclusions:The main contribution of this paper is to use data to mine the principles of the treatment of COVID-19 from the pharmacology of these prescriptions,and the results can be provided theoretical reference for medical workers.
